This Small Business Innovation Research (SBIR) Phase I project will address the challenge of integrating two powerful single-cell analysis tools with the aim of developing and validating new biomarkers for malignancy. The deformability of invasive cells has long been hypothesized to confer their ability to migrate through tight tissue barriers and form metastases. Recently, this idea has been supported by mechanical measurements of cells either isolated from or directly in biological fluid specimens. This convergence of ideas from both biological and physical sciences represents a mechanical biomarker, and tools to be employed clinically to assay these properties are rapidly being developed. Cytovale?s technology measures cell deformability at a throughput of several thousand cells per second, comparable to the ubiquitous flow cytometer, which allows immediate measurement of cells directly in biological fluids. This technology has a demonstrated utility: highly sensitive detection of malignancy in cellularly heterogeneous clinical pleural effusions. Its integration with fluorescence in this project will provide a transformational research and clinical tool, well-aligned with the critical aims of improving patient care and reducing costs through automation, early detection of disease, and use of quantitative, novel biomarkers.
The broader impact/commercial potential of this project is realized by appreciating the applicability of the technology across research and clinical settings. Even without integration with fluorescence (flow) cytometry the technology has demonstrated its utility as a sensitive detector of malignancy in clinical specimens, specifically, pleural effusions. However, cell mechanics is an attractive biomarker for invasiveness, and is likely conserved throughout cells found in many biological fluids, including urine and fine needle aspirates. The proposed activity will further enhance the technology?s diagnostic accuracy. The instruments developed by Cytovale will be placed in clinical cytology labs to complement gold standard cytological methods, performing high sensitivity screens of biological fluids and eliminating unnecessary, invasive, and costly follow-up procedures. The hybrid instrument will also be an especially powerful tool for exploring connections between cell mechanics and traditional markers, which greatly extends the number of research laboratories which would benefit from this enabling technology.
Many disease states significantly alter cells’ mechanical properties through changes in nuclear and cytoskeletal structure. Using innovative microfluidic technology, CytoVale quantifies these changes, providing a rapid, low-cost method of detecting disease. This cytometry platform has been academically validated with clinical samples, cell lines, and disease models. However, prior to this project our technology was primarily operable only by a handful of expert users. This NSF Phase I award enabled us to take this breadboard instrument and develop a powerful, user-friendly instrument while integrating more conventional fluorescence measurements. By integrating conventional flow cytometry-style measurements with this platform, and developing a robust, saleable instrument we have achieved a research-grade instrument, expanding access to the technology to early adopters and furthering our commercial pursuit of clinical diagnostics applications. These measurements have been shown to exquisitely distinguish between cell states (i.e., benign vs. malignant epithelial cells, pluripotent vs. differentiated stem cells, and resting vs. activated immune cells) and have been demonstrated to provide diagnostically-relevant information. Exploiting the system’s fast turnaround time, and building off of recent in vitro and animal model data showing distinct mechanical signatures of white blood cell (WBC) activation, CytoVale has identified sepsis diagnosis as a compelling commercial application. Sepsis, an uncontrolled systemic response to local infection by bacteria or fungi, is responsible for more deaths than prostate cancer, breast cancer, and AIDS combined and is associated with ~$17B in annual U.S. healthcare expenditures. We anticipate that providing emergency department physicians with an earlier diagnostic will profoundly influence clinical outcomes (currently ~40% mortality), costs (>$22,000/case), and the quality of life for survivors and their families. Beyond the adult sepsis screening market, CytoVale has identified several additional indications including an adjacent neonatal sepsis, a bladder cancer detection, academic research tools, and drug development. The products of this award will enable exploration of previously untapped biomarkers for these critical diagnostic needs.
